Development of potency assays for a plasmid containing vascular endothelial growth factor 2

We have developed analytical methods to measure the biological functions of pVGI.1(VEGF2), a naked plasmid DNA product containing vascular endothelial growth factor 2 used in clinical trials for coronary artery diseases (CAD) and peripheral artery diseases (PAD). After being injected into muscles, v...

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Autores principales:	Huang,Li-chun, Chin,Emily, Chiang,Yawen L
Lenguaje:	English
Publicado:	Pontificia Universidad Católica de Valparaíso 2010
Materias:	cell proliferation plasmid DNA potency assays receptor binding VEGF-2
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582010000100001
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spelling	oai:scielo:S0717-345820100001000012010-04-28Development of potency assays for a plasmid containing vascular endothelial growth factor 2Huang,Li-chunChin,EmilyChiang,Yawen L cell proliferation plasmid DNA potency assays receptor binding VEGF-2 We have developed analytical methods to measure the biological functions of pVGI.1(VEGF2), a naked plasmid DNA product containing vascular endothelial growth factor 2 used in clinical trials for coronary artery diseases (CAD) and peripheral artery diseases (PAD). After being injected into muscles, vascular endothelial growth factor 2 (VEGF-2), presumably expressed in muscle tissues, binds to the endothelial cell receptors VEGFR2 or VEGFR3, triggering the downstream responses including cell proliferation and vascularization. As it is important to make sure clinical material is biological active, we developed a quantitative assay first to measure the receptor binding activity of the pVGI.1(VEGF2) gene product expressed by the transfected host cells, and then a qualitative assay to confirm the cell proliferation promoting activity of the expressed protein. In both assays the signals were plotted directly against input DNA concentrations used to transfect the host cells. We confirmed specificity for both assays. In addition, we demonstrated acceptable levels of spike recovery (86.7-116%), precision (largest relative standard deviation (RSD)=19.3%), linearity and range (60-140% relative potency, 15 - 35 µg/mL) for the quantitative assay. We intend to use the potency assays for routine lot release and stability studies.info:eu-repo/semantics/openAccessPontificia Universidad Católica de ValparaísoElectronic Journal of Biotechnology v.13 n.1 20102010-01-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582010000100001en10.4067/S0717-34582010000100001
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	cell proliferation plasmid DNA potency assays receptor binding VEGF-2
spellingShingle	cell proliferation plasmid DNA potency assays receptor binding VEGF-2 Huang,Li-chun Chin,Emily Chiang,Yawen L Development of potency assays for a plasmid containing vascular endothelial growth factor 2
description	We have developed analytical methods to measure the biological functions of pVGI.1(VEGF2), a naked plasmid DNA product containing vascular endothelial growth factor 2 used in clinical trials for coronary artery diseases (CAD) and peripheral artery diseases (PAD). After being injected into muscles, vascular endothelial growth factor 2 (VEGF-2), presumably expressed in muscle tissues, binds to the endothelial cell receptors VEGFR2 or VEGFR3, triggering the downstream responses including cell proliferation and vascularization. As it is important to make sure clinical material is biological active, we developed a quantitative assay first to measure the receptor binding activity of the pVGI.1(VEGF2) gene product expressed by the transfected host cells, and then a qualitative assay to confirm the cell proliferation promoting activity of the expressed protein. In both assays the signals were plotted directly against input DNA concentrations used to transfect the host cells. We confirmed specificity for both assays. In addition, we demonstrated acceptable levels of spike recovery (86.7-116%), precision (largest relative standard deviation (RSD)=19.3%), linearity and range (60-140% relative potency, 15 - 35 µg/mL) for the quantitative assay. We intend to use the potency assays for routine lot release and stability studies.
author	Huang,Li-chun Chin,Emily Chiang,Yawen L
author_facet	Huang,Li-chun Chin,Emily Chiang,Yawen L
author_sort	Huang,Li-chun
title	Development of potency assays for a plasmid containing vascular endothelial growth factor 2
title_short	Development of potency assays for a plasmid containing vascular endothelial growth factor 2
title_full	Development of potency assays for a plasmid containing vascular endothelial growth factor 2
title_fullStr	Development of potency assays for a plasmid containing vascular endothelial growth factor 2
title_full_unstemmed	Development of potency assays for a plasmid containing vascular endothelial growth factor 2
title_sort	development of potency assays for a plasmid containing vascular endothelial growth factor 2
publisher	Pontificia Universidad Católica de Valparaíso
publishDate	2010
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582010000100001
work_keys_str_mv	AT huanglichun developmentofpotencyassaysforaplasmidcontainingvascularendothelialgrowthfactor2 AT chinemily developmentofpotencyassaysforaplasmidcontainingvascularendothelialgrowthfactor2 AT chiangyawenl developmentofpotencyassaysforaplasmidcontainingvascularendothelialgrowthfactor2
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Development of potency assays for a plasmid containing vascular endothelial growth factor 2

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